Method of treating conditions associated with intestinal ischemia/reperfusion

ABSTRACT

The present invention provides methods of treating adverse physiological effects associated with intestinal ischemia/reperfusion by administering to a subject suffering from the effects of intestinal ischemia/reperfusion an effective amount of a BPI protein product.

BACKGROUND OF THE INVENTION

[0001] The present invention relates to therapeutic uses ofbactericidal/permeability-increasing (BPI) protein products for thetreatment of adverse physiological effects associated with intestinalischemia/reperfusion.

[0002] Reperfusion of ischemic intestines is associated with profoundcardiovascular and respiratory dysfunction that may lead to shock anddeath. A variety of mediators are believed to be released from theischemic tissue that could lead to cardiorespiratory collapse, includingoxygen free radicals, protanoids, and platelet activating factor.

[0003] During ischemia, breakdown of the intestinal mucosal permeabilitybarrier may result in translocation of endotoxin and/or bacteria fromthe intestinal lumen. Endotoxin has been detected in the portal veinafter intestinal ischemia/reperfusion. However, a role for translocatedbacteria or endotoxin in intestinal ischemia/reperfusion injury has notbeen clearly defined.

[0004] Bactericidal/permeability-increasing protein (BPI) is a proteinisolated from the granules of mammalian PMNs, which are blood cellsessential in the defense against invading microorganisms. Human BPIprotein has been isolated from polymorphonuclear neutrophils by acidextraction combined with either ion exchange chromatography [Elsbach, J.Biol. Chem., 254:11000 (1979)] or E. coli affinity chromatography[Weiss, et al., Blood, 69:652 (1987)] referred to herein as natural BPIand has potent bactericidal activity against a broad spectrum ofgram-negative bacteria. The molecular weight of human BPI isapproximately 55,000 daltons (55 kD). The amino acid sequence of theentire human BPI protein, as well as the DNA encoding the protein, havebeen elucidated in FIG. 1 of Gray et al., J. Biol. Chem., 264:9505(1989), incorporated herein by reference.

[0005] The bactericidal effect of BPI has been shown to be highlyspecific to sensitive gram-negative species, while non-toxic for othermicroorganisms and for eukaryotic cells. The precise mechanism by whichBPI kills bacteria is as yet unknown, but it is known that BPI mustfirst attach to the surface of susceptible gram-negative bacteria. Thisinitial binding of BPI to the bacteria involves electrostaticinteractions between the basic BPI protein and the negatively chargedsites on lipopolysaccharides (LPS). LPS has been referred to as“endotoxin” because of the potent inflammatory response that itstimulates. LPS induces the release of mediators by host inflammatorycells which may ultimately result in irreversible endotoxic shock. BPIbinds to Lipid A, the most toxic and most biologically active componentof LPS.

[0006] In susceptible bacteria, BPI binding is thought to disrupt LPSstructure, leading to activation of bacterial enzymes that degradephospholipids and peptidoglycans, altering the permeability of thecell's outer membrane, and ultimately causing cell death by an as yetunknown mechanism. BPI is also capable of neutralizing the endotoxicproperties of LPS to which it binds. Because of its gram-negativebactericidal properties and its ability to neutralize LPS, BPI can beutilized for the treatment of mammals suffering from diseases caused bygram-negative bacteria, such as bacteremia or sepsis. Bahrami et al.,Int'l Endotoxin Soc. Meeting, Vienna, Austria (August 1992), disclosethe use of a BPI protein for the treatment of hemorrhagic shock.

[0007] A proteolytic fragment corresponding to the N-terminal portion ofhuman BPI holoprotein possesses the antibacterial efficacy of thenaturally-derived 55 kD human holoprotein. In contrast to the N-terminalportion, the C-terminal region of the isolated human BPI proteindisplays only slightly detectable anti-bacterial activity. Ooi, et al.,J. Exp. Med., 174:649 (1991). A BPI N-terminal fragment, comprisingapproximately the first 199 amino acid residues of the human BPIholoprotein and referred to as “rBPI₂₃”, has been produced byrecombinant means as a 23 kD protein. Gazzano-Santoro et al., Infect.Immun. 60:4754-4761 (1992).

SUMMARY OF THE INVENTION

[0008] The present invention provides novel methods for the treatment ofadverse physiological effects associated with intestinalischemia/reperfusion comprising administering BPI protein products to asubject suffering from the effects of intestinal ischemia/reperfusion.Specifically, the invention provides methods of treating the adversephysiological effects, including cardiac and hemodynamic effects, ofintestinal ischemia/reperfusion resulting from a variety of causes. Suchcauses include mesenteric artery ischemia which is secondary toocclusions resulting from atherosclerosis, embolisms or arterial spasms;ischemia resulting from vessel occlusions in other segments of thebowel; ischemic colitis and intestinal torsion such as occurs in infantsand particularly in animals. In particular, the invention providesmethods for treating the adverse cardiac and other effects of intestinalischemia and reperfusion associated with myocardial infarction.

[0009] The invention thus provides methods for treatment of sepsis-likeconditions associated with intestinal ischemia/reperfusion comprisingadministering to a subject an amount of a BPI protein product effectiveto alleviate adverse physiological effects resulting from the presenceof bacteria, bacterial particulates and endotoxin present in the bodyand in circulation in the blood.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010]FIG. 1 depicts the hemodynamic effects of intestinal reperfusion;

[0011]FIG. 2 depicts the effects of a BPI protein product onreperfusion-induced hemodynamic dysfunction;

[0012]FIGS. 3a and 3 b depict the effects of a BPI protein product onhypotension resulting from intestinal ischemia/reperfusion;

[0013]FIGS. 4a and 4 b depict the effects of a BPI protein product onbradycardia resulting from intestinal ischemia/reperfusion;

[0014]FIG. 5 depicts the effects of a BPI protein product on respiratorydepression resulting from intestinal ischemia/reperfusion;

[0015]FIG. 6 depicts the effects of a BPI protein product on arrythmiasresulting from intestinal ischemia/reperfusion;

[0016]FIG. 7 depicts the effects of a BPI protein product on thesurvival time for rats subjected to intestinal ischemia/reperfusion; and

[0017]FIGS. 8a and 8 b depict the quantity of bacteria isolated fromtissues of rats subjected to intestinal ischemia/reperfusion and thenumber of rats in which the bacteria were detected.

DETAILED DESCRIPTION OF THE INVENTION

[0018] The invention provides methods of treatment of the adverseeffects of intestinal ischemia/reperfusion by administering BPI proteinproducts to subjects suffering from the effects of intestinal ischemiaand reperfusion. According to one aspect of the invention, the adversecardiac and hemodynamic effects including cardiac depression, arrthymiaand hypotension associated with intestinal ischemia/reperfusion arealleviated by administration of effective amounts of BPI proteinproducts. In particular, because these studies demonstrate the adversecardiac and hemodynamic effects of intestinal ischemia/reperfusion, theadministration of BPI protein products as an adjunctive therapy for thetreatment of myocardial infarction would be particularly useful. The BPIprotein products are preferably administered systemically such asintravenously, or by intramuscular or subcutaneous injection.

[0019] As used herein, “BPI protein product” includes naturally andrecombinantly produced bactericidal/permeability-increasing protein;natural, synthetic, and recombinant biologically active polypeptidefragments of bactericidal/permeability-increasing protein; andbiologically active polypeptide analogs or variants including hybridfusion proteins, of either bactericidal/permeability-increasing proteinor biologically active fragments thereof. The BPI protein productsincluding biologically active fragments of BPI holoprotein which are tobe administered according to the methods of this invention may begenerated and/or isolated by any means known in the art. U.S. Pat. No.5,198,541, the disclosure of which is hereby incorporated by reference,discloses recombinant genes encoding and methods for expression of BPIproteins. Co-owned, copending U.S. patent application Ser. No.07/885,501 and a continuation-in-part thereof, U.S. patent applicationSer. No. 08/072,063 filed May 19, 1993, which are hereby incorporated byreference, disclose novel methods for the purification of recombinantBPI protein products expressed in and secreted from geneticallytransformed mammalian host cells in culture, and discloses how one mayproduce large quantities of recombinant BPI products suitable forincorporation into stable, homogeneous pharmaceutical preparations.

[0020] Biologically active fragments of BPI include biologically activemolecules that contain the same amino acid sequence as a BPIholoprotein, except that the molecule lacks amino-terminal amino acids,internal amino acids, and/or carboxy-terminal amino acids of theholoprotein. Amino-terminal fragments of BPI comprising up to about thefirst 200 amino acid residues of BPI are contemplated as beingparticularly useful according to the invention. By way of nonlimitingexamples, such fragments include those described herein and a natural 25Kd fragment and a recombinant 23 Kd, 199 amino acid residueamino-terminal fragment of the human BPI holoprotein referred to asrBPI₂₃. See, Gazzano-Santoro et al., Infect. Immun. 60:4754-4761 (1992).In that publication, an expression vector was used as a source of DNAencoding a recombinant expression product (rBPI₂₃) having the 31-residuesignal sequence and the first 199 amino acids of the N-terminus of themature human BPI, as set out in SEQ ID NOS: 1 and 2 taken from Gray etal., supra, except that valine at position 151 is specified by GTGrather than GTC and residue 185 is glutamic acid (specified by GAG)rather than lysine (specified by AAG). Recombinant holoprotein referredto herein as rBPI has also been produced having the sequence set out inSEQ ID NOS: 1 and 2 taken from Gray et al., supra, with the exceptionsnoted for rBPI₂₃.

[0021] Biologically active analogs and variants of BPI include, but arenot limited to, recombinant hybrid fusion proteins comprising BPIholoprotein or biologically active fragment thereof, and at least aportion of at least one other polypeptide. Such proteins are describedby Theofan et al. in co-owned, copending U.S. patent application Ser.No. 07/885,911, and a continuation-in-part application thereof U.S.patent application Ser. No. 08/064693 filed May 19, 1993, which areincorporated herein by reference in their entirety and include hybridfusion proteins comprising, at the amino terminal end, a BPI protein ora biologically active fragment thereof and, at the carboxy terminal end,at least one constant domain of an immunoglobulin heavy chain or allelicvariant thereof.

[0022] Biologically active analogs and variants of BPI also include, butare not limited to, BPI protein products wherein one or more amino acidresidues have been replaced by a different amino acid. For example,co-owned, copending U.S. patent application Ser. No. 08/013,801 (Theofanet al., “Stable Bactericidal/Permeability-Increasing Protein Productsand Pharmaceutical Compositions Containing the Same,” filed Feb. 2,1993) which is incorporated herein by reference, discloses polypeptideanalogs of BPI and BPI fragments wherein a cysteine residue at position132 or at position 135 is replaced by a different amino acid. Apreferred BPI protein product described by this application comprisesthe first 199 amino acids of BPI holoprotein but wherein the cysteine atresidue number 132 is substituted with alanine and is designatedrBPI₂₁Δcys.

[0023] The administration of BPI protein products is preferablyaccomplished with a pharmaceutical composition comprising a BPI proteinproduct and a pharmaceutically acceptable diluent, adjuvant, or carrier.The BPI protein product composition may be administered without or inconjunction with known surfactants, other chemotherapeutic agents oradditional known antibiotics. A preferred pharmaceutical compositioncontaining BPI protein products comprises BPI at a concentration ofabout 1 to 2 mg/ml in citrate buffered saline (0.02 M citrate, 0.15 MNaCl, pH 5.0) comprising 0.1% by weight of poloxamer 188 (Pluronic F-68,BASF Wyandotte, Parsippany, N.J.) and 0.002% by weight of polysorbate 80(Tween 80, ICI Americas Inc., Wilmington, Del.). Such preferredcombinations are described in co-owned, copending, U.S. patentapplication Ser. No. 08/190,869 filed Feb. 2, 1994 which is acontinuation in part of U.S patent application Ser. No. 08/012,360(McGregor et al., “Improved Pharmaceutical Composition” filed Feb. 2,1993), the disclosures of which are incorporated herein by reference.

[0024] Other BPI protein products useful according to the methods of theinvention are BPI peptides such as those described in co-owned andcopending U.S. patent application Ser. No. 08/209,762 filed Mar. 11,1994 which is a continuation-in-part of U.S. patent application Ser. No.08/183,222 filed Jan. 14, 1994 which is a continuation-in-part of U.S.patent application Ser. No. 08/093,202 filed Jul. 15, 1993 which is acontinuation in part of U.S. patent application Ser. No. 08/030,644filed Mar. 12, 1993 the disclosures of which are hereby incorporated byreference.

[0025] Practice of the methods of the present invention is illustratedin the following examples wherein: Example 1 discloses the effect ofadministration of a BPI protein product on blood pressure, heart rateand respiratory rate of rats subjected to intestinalischemia/reperfusion. Example 2 discloses the effect of administrationof a BPI protein product on the translocation of bacteria in ratssubjected to intestinal ischemia/reperfusion.

EXAMPLE 1

[0026] A rat surgical model was used to evaluate the effects of BPIprotein products on the physiological effects associated with intestinalischemia/reperfusion. Specifically Sprague Dawley rats were anesthetizedwith a mixture of 80 mg/kg of ketamine and 4 mg/kg of xylazineadministered by intraperitoneal injection. After a surgical plane ofanesthesia was obtained, a tracheotomy was performed and a trachealcannula was inserted. The animals breathed on their own. A catheter,made of polyethylene tubing, was placed in a femoral artery. Thecatheter was connected to a pressure transducer in order to measureblood pressure. Another catheter was placed in a femoral vein andconnected to an infusion pump.

[0027] The abdominal contents were then exposed via a midline abdominalincision. The superior mesenteric artery (SMA) was visualized at itsjunction with the abdominal aorta and a silk ligature was threadedaround the SMA after it had been gently loosened from the surroundingconnective tissue. The loose ends of the ligature were placed outsidethe animal and the abdominal incision was then closed with surgicalstaples.

[0028] After surgery, the cardiac indices of blood pressure and heartrate were electronically recorded by measurement at the femoral arteryand respiration rate was determined by visual observation for a periodof about 30 to 45 minutes so that all recorded variables were stable.The ligature around the SMA was then tightened by until the SMA wasoccluded. The SMA remained occluded for 90 minutes at which time theligature was loosened to allow reperfusion. Sixty minutes after the SMAwas occluded 7 rats received an intravenous bolus injection ofrBPI₂₁Δcys in a vehicle comprising citrate buffered saline (0.02 Mcitrate, 0.15 M NaCl, pH 5.0) comprising 0.1% by weight of poloxamer 188and 0.002% by weight of polysorbate 80 followed by a constant infusionof 2 mg/kg/hr. Seven other control rats received equal volumes ofvehicle. The infusions continued until death.

[0029] Typical blood pressure and heart rate records for individualuntreated and BPI treated rats are presented in FIGS. 1 and 2respectively. Opening the SMA occlusion after 90 minutes resulted inrapid declines in blood pressure and heart rate of all rats treated withvehicle. Within a few minutes, the heart rate of all but one control ratbegan to oscillate, partially in phase with respiration, but also in aslower, more irregular pattern. In 5 of the 7 control rats thereappeared to be missed beats which are presumably the result ofarrhythmias. In contrast, irregularities of heart rate and arrhythmiaswere seldom observed in rBPI₂₁Δcys treated rats. Administration ofrBPI₂₁Δcys did have an effect in reducing the hypotension resulting fromintestinal ischemia/reperfusion as shown by the results in FIGS. 3a and3 b where FIG. 3a shows the results for both the ischemic andreperfusion phases of the experiment and FIG. 3b shows only the periodof reperfusion. Data are shown for the first 30 minutes after openingthe occlusion because all vehicle-treated rats were dead within 45minutes and all BPI treated rats were dead within 60 minutes.

[0030] The results illustrated in FIGS. 4a and 4 b (where FIG. 4a showsthe results for both the ischemic and reperfusion phases of theexperiment and FIG. 4b shows only the period of reperfusion) show thatthe administration of rBPI₂₁Δcys had the effect of preventingbradycardia resulting from intestinal ischemia/reperfusion.

[0031] The results illustrated in FIG. 5 measuring respiratory rate showthat the administration of the BPI protein product reduces respiratorydepression resulting from the intestinal ischemia/reperfusion injury.The figure only presents data following reperfusion because therespiration rates for vehicle and BPI treated rats were not differentprior to opening the occlusion.

[0032] The effect of intestinal ischemia and reperfusion on rats treatedand untreated with the BPI protein product are set out in Table I belowwhich illustrates the data of FIGS. 3a, 3 b, 4 a, 4 b and 5 where t=0 isimmediately before reperfusion and t=30 is after 30 minutes ofreperfusion.

[0033] The results illustrated in FIG. 6 relating to arrythmia durationshow that administration of the BPI protein product reduces the durationof heart rate irregularities resulting from intestinal reperfusion. Forthis analysis the period of time during which obvious heart rateoscillations or arrhythmias were observed was determined for each ratand then averaged. The results were statistically significant withp<0.001.

[0034] The results illustrated in FIG. 7 show that treatment with theBPI protein product increases survival time in rats suffering fromintestinal ischemia/reperfusion injury (p<0.05). According to thisaspect of the experiment, the time from opening the occlusion untildeath was recorded for each rat. In all cases death was immediatelypreceded by a rapid decline in respiration rate and reduction in tidalvolume. TABLE I Blood Pressure Heat Rate Respiratory (mmHg) (per min.)Rate (per min.) t = 0 t = 30 t = 0 t = 30 t = 0 t = 30 Vehicle 96 ± 5 37± 3 288 ± 2 241 ± 16  51 ± 3 29 ± 3  rBPI₂₁Δcys 96 ± 5 43 ± 5 284 ± 7325 ± 9** 56 ± 4 40 ± 4*

EXAMPLE 2

[0035] In an additional experiment with the rat surgical model ofexperiment 1 two groups of five rats each were surgically prepared andadministered with either rBPI₂₁Δcys, or vehicle in the same manner anddosages as in example 1 except that a blood sample was obtained justprior to death. In addition a third group of five rats was subjected toa sham operation wherein all the surgical procedures were reproducedwith the exception that the SMA was never occluded. After death, samplesof tissue were obtained from the liver, spleen, and mesenteric lymphnodes. The blood was plated on trypticase soy agar and incubatedovernight at 37° C. The tissue samples were then weighed and homogenizedand were similarly plated and incubated overnight at 37° C. The next daythe number of colonies on the plates were counted visually. The numberof bacteria per gram of tissue of each organ is shown in FIG. 8a. Thenumber of rats in which bacteria were detected is shown in FIG. 8b.These results including the sham experiment show that intestinalischemia/reperfusion results in translocation of bacteria, most likelyfrom the gut. Further, the results show that the administration of theBPI protein product reduced the translocation of bacteria resulting fromintestinal ischemia/reperfusion. Analysis of the blood samples indicatedno presence of bacteremia in any of the subject animals.

[0036] Numerous modifications and variations in the practice of theinvention are expected to occur to those skilled in the art uponconsideration of the foregoing description of the presently preferredembodiments thereof. Consequently, the only limitations which should beplaced upon the scope of the present invention are those which appear inthe appended claims.

1 2 1813 base pairs nucleic acid single linear cDNA CDS 31..1491mat_peptide 124..1491 misc_feature “rBPI” 1 CAGGCCTTGA GGTTTTGGCAGCTCTGGAGG ATG AGA GAG AAC ATG GCC AGG GGC 54 Met Arg Glu Asn Met AlaArg Gly -31 -30 -25 CCT TGC AAC GCG CCG AGA TGG GTG TCC CTG ATG GTG CTCGTC GCC ATA 102 Pro Cys Asn Ala Pro Arg Trp Val Ser Leu Met Val Leu ValAla Ile -20 -15 -10 GGC ACC GCC GTG ACA GCG GCC GTC AAC CCT GGC GTC GTGGTC AGG ATC 150 Gly Thr Ala Val Thr Ala Ala Val Asn Pro Gly Val Val ValArg Ile -5 1 5 TCC CAG AAG GGC CTG GAC TAC GCC AGC CAG CAG GGG ACG GCCGCT CTG 198 Ser Gln Lys Gly Leu Asp Tyr Ala Ser Gln Gln Gly Thr Ala AlaLeu 10 15 20 25 CAG AAG GAG CTG AAG AGG ATC AAG ATT CCT GAC TAC TCA GACAGC TTT 246 Gln Lys Glu Leu Lys Arg Ile Lys Ile Pro Asp Tyr Ser Asp SerPhe 30 35 40 AAG ATC AAG CAT CTT GGG AAG GGG CAT TAT AGC TTC TAC AGC ATGGAC 294 Lys Ile Lys His Leu Gly Lys Gly His Tyr Ser Phe Tyr Ser Met Asp45 50 55 ATC CGT GAA TTC CAG CTT CCC AGT TCC CAG ATA AGC ATG GTG CCC AAT342 Ile Arg Glu Phe Gln Leu Pro Ser Ser Gln Ile Ser Met Val Pro Asn 6065 70 GTG GGC CTT AAG TTC TCC ATC AGC AAC GCC AAT ATC AAG ATC AGC GGG390 Val Gly Leu Lys Phe Ser Ile Ser Asn Ala Asn Ile Lys Ile Ser Gly 7580 85 AAA TGG AAG GCA CAA AAG AGA TTC TTA AAA ATG AGC GGC AAT TTT GAC438 Lys Trp Lys Ala Gln Lys Arg Phe Leu Lys Met Ser Gly Asn Phe Asp 9095 100 105 CTG AGC ATA GAA GGC ATG TCC ATT TCG GCT GAT CTG AAG CTG GGCAGT 486 Leu Ser Ile Glu Gly Met Ser Ile Ser Ala Asp Leu Lys Leu Gly Ser110 115 120 AAC CCC ACG TCA GGC AAG CCC ACC ATC ACC TGC TCC AGC TGC AGCAGC 534 Asn Pro Thr Ser Gly Lys Pro Thr Ile Thr Cys Ser Ser Cys Ser Ser125 130 135 CAC ATC AAC AGT GTC CAC GTG CAC ATC TCA AAG AGC AAA GTC GGGTGG 582 His Ile Asn Ser Val His Val His Ile Ser Lys Ser Lys Val Gly Trp140 145 150 CTG ATC CAA CTC TTC CAC AAA AAA ATT GAG TCT GCG CTT CGA AACAAG 630 Leu Ile Gln Leu Phe His Lys Lys Ile Glu Ser Ala Leu Arg Asn Lys155 160 165 ATG AAC AGC CAG GTC TGC GAG AAA GTG ACC AAT TCT GTA TCC TCCAAG 678 Met Asn Ser Gln Val Cys Glu Lys Val Thr Asn Ser Val Ser Ser Lys170 175 180 185 CTG CAA CCT TAT TTC CAG ACT CTG CCA GTA ATG ACC AAA ATAGAT TCT 726 Leu Gln Pro Tyr Phe Gln Thr Leu Pro Val Met Thr Lys Ile AspSer 190 195 200 GTG GCT GGA ATC AAC TAT GGT CTG GTG GCA CCT CCA GCA ACCACG GCT 774 Val Ala Gly Ile Asn Tyr Gly Leu Val Ala Pro Pro Ala Thr ThrAla 205 210 215 GAG ACC CTG GAT GTA CAG ATG AAG GGG GAG TTT TAC AGT GAGAAC CAC 822 Glu Thr Leu Asp Val Gln Met Lys Gly Glu Phe Tyr Ser Glu AsnHis 220 225 230 CAC AAT CCA CCT CCC TTT GCT CCA CCA GTG ATG GAG TTT CCCGCT GCC 870 His Asn Pro Pro Pro Phe Ala Pro Pro Val Met Glu Phe Pro AlaAla 235 240 245 CAT GAC CGC ATG GTA TAC CTG GGC CTC TCA GAC TAC TTC TTCAAC ACA 918 His Asp Arg Met Val Tyr Leu Gly Leu Ser Asp Tyr Phe Phe AsnThr 250 255 260 265 GCC GGG CTT GTA TAC CAA GAG GCT GGG GTC TTG AAG ATGACC CTT AGA 966 Ala Gly Leu Val Tyr Gln Glu Ala Gly Val Leu Lys Met ThrLeu Arg 270 275 280 GAT GAC ATG ATT CCA AAG GAG TCC AAA TTT CGA CTG ACAACC AAG TTC 1014 Asp Asp Met Ile Pro Lys Glu Ser Lys Phe Arg Leu Thr ThrLys Phe 285 290 295 TTT GGA ACC TTC CTA CCT GAG GTG GCC AAG AAG TTT CCCAAC ATG AAG 1062 Phe Gly Thr Phe Leu Pro Glu Val Ala Lys Lys Phe Pro AsnMet Lys 300 305 310 ATA CAG ATC CAT GTC TCA GCC TCC ACC CCG CCA CAC CTGTCT GTG CAG 1110 Ile Gln Ile His Val Ser Ala Ser Thr Pro Pro His Leu SerVal Gln 315 320 325 CCC ACC GGC CTT ACC TTC TAC CCT GCC GTG GAT GTC CAGGCC TTT GCC 1158 Pro Thr Gly Leu Thr Phe Tyr Pro Ala Val Asp Val Gln AlaPhe Ala 330 335 340 345 GTC CTC CCC AAC TCC TCC CTG GCT TCC CTC TTC CTGATT GGC ATG CAC 1206 Val Leu Pro Asn Ser Ser Leu Ala Ser Leu Phe Leu IleGly Met His 350 355 360 ACA ACT GGT TCC ATG GAG GTC AGC GCC GAG TCC AACAGG CTT GTT GGA 1254 Thr Thr Gly Ser Met Glu Val Ser Ala Glu Ser Asn ArgLeu Val Gly 365 370 375 GAG CTC AAG CTG GAT AGG CTG CTC CTG GAA CTG AAGCAC TCA AAT ATT 1302 Glu Leu Lys Leu Asp Arg Leu Leu Leu Glu Leu Lys HisSer Asn Ile 380 385 390 GGC CCC TTC CCG GTT GAA TTG CTG CAG GAT ATC ATGAAC TAC ATT GTA 1350 Gly Pro Phe Pro Val Glu Leu Leu Gln Asp Ile Met AsnTyr Ile Val 395 400 405 CCC ATT CTT GTG CTG CCC AGG GTT AAC GAG AAA CTACAG AAA GGC TTC 1398 Pro Ile Leu Val Leu Pro Arg Val Asn Glu Lys Leu GlnLys Gly Phe 410 415 420 425 CCT CTC CCG ACG CCG GCC AGA GTC CAG CTC TACAAC GTA GTG CTT CAG 1446 Pro Leu Pro Thr Pro Ala Arg Val Gln Leu Tyr AsnVal Val Leu Gln 430 435 440 CCT CAC CAG AAC TTC CTG CTG TTC GGT GCA GACGTT GTC TAT AAA 1491 Pro His Gln Asn Phe Leu Leu Phe Gly Ala Asp Val ValTyr Lys 445 450 455 TGAAGGCACC AGGGGTGCCG GGGGCTGTCA GCCGCACCTGTTCCTGATGG GCTGTGGGGC 1551 ACCGGCTGCC TTTCCCCAGG GAATCCTCTC CAGATCTTAACCAAGAGCCC CTTGCAAACT 1611 TCTTCGACTC AGATTCAGAA ATGATCTAAA CACGAGGAAACATTATTCAT TGGAAAAGTG 1671 CATGGTGTGT ATTTTAGGGA TTATGAGCTT CTTTCAAGGGCTAAGGCTGC AGAGATATTT 1731 CCTCCAGGAA TCGTGTTTCA ATTGTAACCA AGAAATTTCCATTTGTGCTT CATGAAAAAA 1791 AACTTCTGGT TTTTTTCATG TG 1813 487 amino acidsamino acid linear protein 2 Met Arg Glu Asn Met Ala Arg Gly Pro Cys AsnAla Pro Arg Trp Val -31 -30 -25 -20 Ser Leu Met Val Leu Val Ala Ile GlyThr Ala Val Thr Ala Ala Val -15 -10 -5 1 Asn Pro Gly Val Val Val Arg IleSer Gln Lys Gly Leu Asp Tyr Ala 5 10 15 Ser Gln Gln Gly Thr Ala Ala LeuGln Lys Glu Leu Lys Arg Ile Lys 20 25 30 Ile Pro Asp Tyr Ser Asp Ser PheLys Ile Lys His Leu Gly Lys Gly 35 40 45 His Tyr Ser Phe Tyr Ser Met AspIle Arg Glu Phe Gln Leu Pro Ser 50 55 60 65 Ser Gln Ile Ser Met Val ProAsn Val Gly Leu Lys Phe Ser Ile Ser 70 75 80 Asn Ala Asn Ile Lys Ile SerGly Lys Trp Lys Ala Gln Lys Arg Phe 85 90 95 Leu Lys Met Ser Gly Asn PheAsp Leu Ser Ile Glu Gly Met Ser Ile 100 105 110 Ser Ala Asp Leu Lys LeuGly Ser Asn Pro Thr Ser Gly Lys Pro Thr 115 120 125 Ile Thr Cys Ser SerCys Ser Ser His Ile Asn Ser Val His Val His 130 135 140 145 Ile Ser LysSer Lys Val Gly Trp Leu Ile Gln Leu Phe His Lys Lys 150 155 160 Ile GluSer Ala Leu Arg Asn Lys Met Asn Ser Gln Val Cys Glu Lys 165 170 175 ValThr Asn Ser Val Ser Ser Lys Leu Gln Pro Tyr Phe Gln Thr Leu 180 185 190Pro Val Met Thr Lys Ile Asp Ser Val Ala Gly Ile Asn Tyr Gly Leu 195 200205 Val Ala Pro Pro Ala Thr Thr Ala Glu Thr Leu Asp Val Gln Met Lys 210215 220 225 Gly Glu Phe Tyr Ser Glu Asn His His Asn Pro Pro Pro Phe AlaPro 230 235 240 Pro Val Met Glu Phe Pro Ala Ala His Asp Arg Met Val TyrLeu Gly 245 250 255 Leu Ser Asp Tyr Phe Phe Asn Thr Ala Gly Leu Val TyrGln Glu Ala 260 265 270 Gly Val Leu Lys Met Thr Leu Arg Asp Asp Met IlePro Lys Glu Ser 275 280 285 Lys Phe Arg Leu Thr Thr Lys Phe Phe Gly ThrPhe Leu Pro Glu Val 290 295 300 305 Ala Lys Lys Phe Pro Asn Met Lys IleGln Ile His Val Ser Ala Ser 310 315 320 Thr Pro Pro His Leu Ser Val GlnPro Thr Gly Leu Thr Phe Tyr Pro 325 330 335 Ala Val Asp Val Gln Ala PheAla Val Leu Pro Asn Ser Ser Leu Ala 340 345 350 Ser Leu Phe Leu Ile GlyMet His Thr Thr Gly Ser Met Glu Val Ser 355 360 365 Ala Glu Ser Asn ArgLeu Val Gly Glu Leu Lys Leu Asp Arg Leu Leu 370 375 380 385 Leu Glu LeuLys His Ser Asn Ile Gly Pro Phe Pro Val Glu Leu Leu 390 395 400 Gln AspIle Met Asn Tyr Ile Val Pro Ile Leu Val Leu Pro Arg Val 405 410 415 AsnGlu Lys Leu Gln Lys Gly Phe Pro Leu Pro Thr Pro Ala Arg Val 420 425 430Gln Leu Tyr Asn Val Val Leu Gln Pro His Gln Asn Phe Leu Leu Phe 435 440445 Gly Ala Asp Val Val Tyr Lys 450 455

What is claimed is:
 1. A method of treating adverse physiologicaleffects associated with intestinal ischemia/reperfusion comprisingadministering to a subject suffering from the effects of intestinalischemia/reperfusion an effective amount of a BPI protein product. 2.The method of claim 1 wherein said intestinal ischemia/reperfusion isassociated with occlusion of an intestinal artery.
 3. The method ofclaim 2 wherein said intestinal ischemia/reperfusion is associated withocclusion of the mesenteric artery.
 4. The method of claim 1 whereinsaid intestinal ischemia/reperfusion is associated with myocardialinfarction.
 5. The method of claim 1 wherein said intestinalischemia/reperfusion is associated with intestinal torsion.
 6. Themethod of claim 1 wherein said adverse physiological effects are cardiaceffects.
 7. The method of claim 1 wherein said adverse physiologicaleffects are hemodynamic effects.
 8. The method of claim 1 wherein theBPI protein product is an amino-terminal fragment of BPI.
 9. The methodof claim 1 wherein the BPI protein product is rBPI₂₁Δcys.
 10. The methodof claim 1 wherein the BPI protein product is administered inconjunction with a pharmaceutically-acceptable diluent, adjuvant orcarrier.